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Utility of dynamic testosterone profiling for assessing human hepatic culture systems for 3D tissue engineering/pharma applications

Nelson, L. N. and Grant, H. G. and Henderson, C. H. and Navarro, M. N. and Plevris, J. P. and Treskes, P. T. (2013) Utility of dynamic testosterone profiling for assessing human hepatic culture systems for 3D tissue engineering/pharma applications. International Journal of Artificial Organs, 36 (8). p. 586. ISSN 0391-3988

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Objectives: The generation of an in vitro 3D organotypic human hepatic model as a surrogate for primary tissue and a platform for drug testing requires various configurations and dimensions of culture to be tested in regard to their biotransformation capabilities. We tested a non-invasive assay for dynamic functional profiling of CYP3A4 activity as a potential tool for the functional assessment of culture formats.Methods & Materials: Human hepatic cell lines [C3A or HepaRG] grown on uncoated or collagen-coated 6-well plates were induced for 24 h with rifampicinand subsequently exposed to 50 μM testosterone for 2 h, before samples were taken and measured via HPLC. Results: HepaRG cells displayed a significantly higher relative turnover of testosterone metabolites (56.47±7.60%, p<0.0001) than both C3A culture formats (Collagen-coated: 1.95±0.19%, uncoated: 2.11±0.45%). Interestingly, the test also indicated more intact metabolic pathways in HepaRGs as shown by the relative abundance of various metabolites, whereas C3A samples only contained androstenedione or in the case of C3As grown on collagen-coated dishes, low additional amounts of 16α-hydroxytestosterone (11.73 vs. 88.27±7.42%). Conclusions: The dynamic testosterone profiling assay gives insight into the relative CYP3A4 specific metabolic activity of hepatic cell lines in vitro. As such,the assay is an attractive tool for the non-invasive assessment of metabolic activity of engineered tissue constructs and the influence of extracellular matrix composition on functionality in comparison to primary tissue and is currently being applied to CYP1A2-specific metabolism.